title: Sporozoite proteome analysis of Cryptosporidium parvum by one-dimensional SDS-PAGE and liquid chromatography tandem mass spectrometry.
abstract: Despite the development of new technologies, new challenges still remain for large scale proteomic profiling when dealing with complex biological mixtures. Fractionation prior to liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis is usually the preferred method to reduce the complexity of any biological sample. In this study, a gel LC-MS/MS approach was used to explore the stage specific proteome of Cryptosporidium (C.) parvum. To accomplish this, the sporozoite protein of C. parvum was first fractionated using SDS-PAGE with subsequent LC-MS/MS analysis. A total of 135 protein hits were recorded from 20 gel slices (from same gel lane), with many hits occurring in more than one band. Excluding all non-Cryptosporidium entries and proteins with multiple hits, 33 separate C. parvum entries were identified during the study. The overall goal of this study was to reduce sample complexity by protein fractionation and increase the possibility of detecting proteins present in lower abundance in a complex protein mixture.
authors.affiliation: Department of Veterinary Preclinical Science, Faculty of Veterinary Science, University of Liverpool, Liverpool L69 7ZJ, UK. zsiddiki@gmail.com
authors.forename: Amam Zonaed
authors.initials: AZ
authors.lastname: Siddiki
mesh_terms: D005591:Chemical Fractionation / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods / Q000662:veterinary; D016785:Cryptosporidium parvum / Q000737:chemistry* / Q000254:growth & development / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods / Q000662:veterinary; D020869:Gene Expression Profiling / Q000379:methods* / Q000662:veterinary; D020543:Proteome / Q000032:analysis; D040901:Proteomics / Q000379:methods*; D015800:Protozoan Proteins / Q000032:analysis*; D034101:Sporozoites / Q000737:chemistry / Q000378:metabolism; D053719:Tandem Mass Spectrometry / Q000379:methods / Q000662:veterinary
chemical_list: D020543:Proteome; D015800:Protozoan Proteins
keywords: Cryptosporidium; SDS-PAGE; bioinformatics; proteomics; tandem mass spectrometry